LI Sheng-li, YU Xiang-dong, WANG Wei-jie and et al. Construction of indirect ELISA method using rabies virus G gene expressed in prokaryotic system[J]. Journal of Light Industry, 2014, 29(4): 24-28. doi: 10.3969/j.issn.2095-476X.2014.04.006
Citation:
LI Sheng-li, YU Xiang-dong, WANG Wei-jie and et al. Construction of indirect ELISA method using rabies virus G gene expressed in prokaryotic system[J]. Journal of Light Industry, 2014, 29(4): 24-28.
doi:
10.3969/j.issn.2095-476X.2014.04.006
Construction of indirect ELISA method using rabies virus G gene expressed in prokaryotic system
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Jiyuan Institute of Animal Health Inspection, Jiyuan 459000, China;
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Luoyang Center for Animal Disease Control and Prevention, Luoyang 471002, China
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Received Date:
2014-04-21
Available Online:
2014-07-15
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Abstract
In order to acquire rabies virus G gene and express it in E.coli and establish the evaluation method for rabies virus immune, according to Rabies virus ERA G gene sequence to design primer sequences,RNA was extracted from rabies virus,the first strand cDNA was synthesized by reverse transcription.G gene fragments were amplified by PCR.Then the gene fragment was cloned to the prokaryotic expression vector pET28a (+) to construct PET-RVG.The positive recombinant plasmids were transfected into E.coli BL21(DE3)gold,with IPTG induced expression and determined the best expression condition.Using Niaffinity chromatography purification with G protein,and the indirect ELISA assay for the detection of rabies virus antibodies in serum was established based on the G protein.The results showed that the purity degree was 96%.
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